Lancemaside A Derived from Codonopsis lanceolata Shoots Modulates the Immunostimulatory Responses by Enhancing TLR4 Signaling Pathway in RAW 264.7 Macrophages

Immune function plays a critical role in defending the body against harmful external stimuli, such as pathogens, and contributes to the maintenance of overall physiological homeostasis. Activation of the immune system is essential for enhancing host defense and preventing immune-related and infectious diseases. In this study, we investigated the immune-enhancing potential of Codonopsis lanceolata shoot extract using the murine macrophage cell line RAW 264.7. Among the solvent extracts tested, the ethanol extract exhibited the highest nitric oxide (NO) production. Treatment with the ethanol extract significantly increased the expression of immune-related genes, including iNOS, COX-2, p-p65, and MAPK. LC-MS/MS analysis identified the major compounds present in the ethanol extract, and in silico docking analysis expected that lancemaside A exhibited the strongest binding affinity to TLR4, a receptor implicated in innate immune signaling. Treatment of RAW 264.7 cells with lancemaside A not only induced the highest NO production but also showed a strong interaction with TLR4. Additionally, lancemaside A enhanced phagocytic activity in a dose-dependent manner and upregulated the expression of immune-related genes. Collectively, these findings suggest that lancemaside A from the ethanol extract of C. lanceolata shoots may serve as a promising functional ingredient for enhancing immune responses and could be developed as a potential immunomodulatory food material.